Invitrogen

ARF6 Polyclonal Antibody

3 Published Figures

6 References

View all (14) ARF6 antibodies

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ARF6 Antibody in Immunofluorescence (IF) — Immunofluorescent analysis of Arf6 (green) in HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 5% normal goat serum (Product # 31873) for 15 minutes at room temperature. Cells were probed with an Arf6 polyclonal antibody (Product # PA1-093) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-rabbit IgG secondary antibody (Product # 35552) at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan at 20X magnification.

ARF6 Antibody in Immunohistochemistry (IHC) — Immunohistochemistry was performed on normal biopsies of deparaffinized human heart tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a Rabbit Polyclonal Antibody recognizing Arf6 (Product # PA1-093) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

ARF6 Antibody in Western Blot (WB) — Western blot analysis of Arf6 was performed by loading 50 µg of various whole cell lysate onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% Milk/TBST for at least 1 hour. Membranes were then probed with a rabbit polyclonal antibody recognizing Arf6 (Product # PA1-093) at a dilution of 1:1000 overnight at 4°C on a rocking platform. Membranes were then washed in TBS-0.1%Tween 20 and probed with a goat anti-rabbit-HRP secondary antibody (Product # 31460) at a dilution of 1:10000 for at least one hour. Membranes were washed and chemiluminescent detection was performed using Super Signal West Pico (Product # 34080).

ARF6 Antibody — Figure 6. The levels of activated Arf6 in cortical tissue are elevated because of Iqsec2 KO. (A) Biochemical assays to measure the levels of activated Arf6 (G-LISA) undertaken in cortical tissues of animals across postnatal development between 2 and 9 mo of age show that (A) the levels of activated Arf6 in wild-type male mice (WT/black) (n = 4) are elevated in age-matched wild-type female mice (WT/grey) (n = 5). (B) Iqsec2 KO hemizygous males (KO/blue, n = 6) and heterozygous KO females (Het/pink; n = 9) both display increased levels of activated Arf6 compared with the sex-matched wild-type controls listed above. (C) The abundance of Arf6 protein measured by immunoblot was not significantly different between any genotype groups. The Iqsec2 KO animals with observed seizures are denoted by stars. Mean (+-SEM) data presented, where * indicates significance between WT female control and HET/KO, P < 0.05, 2-tailed, unpaired t test.

Product Details

Applications

Tested Dilution

Publications

Immunocytochemistry (ICC)

1:20-1:100

View 1 publication 1 publication

Immunofluorescence (IF)

1:20 - 1:100

View 1 publication 1 publication

Immunohistochemistry (Paraffin) (IHC (P))

1:20-100

Western Blot (WB)

1:1000

View 3 publications 3 publications

Miscellaneous PubMed (Misc)

View 1 publication 1 publication

Product Specifications

Species

Human, Mouse, Non-human primate

Published species

Human, Mouse

Expression System

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

C-terminal human recombinant protein fragment

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Protein A

Storage buffer

PBS with 1mg/mL BSA, 30% glycerol

Contains

0.05% sodium azide

Storage conditions

-20° C, Avoid Freeze/Thaw Cycles

RRID

AB_2539860

Target Information

ARF6 (ADP-ribosylation factor 6) is a broadly expressed, differentially regulated GTPase that is present in cytoplasm and on both cell-surface and intracellular membranes and whose functions may include tissue-specific effects on vesicular trafficking during cellular differentiation.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.